In vitro cytotoxicity of asbestos and man-made vitreous fibers: roles of fiber length, diameter and composition

Abstract

The present study investigated (i) the impact of various fiber parameters on in vitro toxicity to cells and (ii) the validity of an in vitro test system as a toxic screen for fibrous materials. Chinese hamster ovary cells were exposed in vitro to a series of size-selected inorganic test fibers that represented a range of different diameters, lengths and compositions (glass, refractory ceramic, mineral wool, asbestos). Toxic end-points included inhibition of proliferation, induction of micronuclei and polynuclei and viability. For all compositions tested, toxic effects were similar: a concentration-dependent decrease in proliferation and increase in incidence of morphologically abnormal nuclei with minor decreases in viability. Diameter-dependent differences in toxicity were slight or absent for fiber diameters ranging from 0.3-7 microns when concentration was expressed as number of fibers/cm2. Length-dependent differences in toxicity were, however, striking. EC50 values (concentration in fibers/cm2 that reduced cell proliferation to 50% of unexposed control cultures) plotted against fiber length produced a hyperbolic curve, demonstrating that toxicity increases with fiber length up to 20 microns. All fibers tested fell on this hyperbola. These data suggest that: (a) the primary toxic effect of fibers on CHO cells is the induction of nuclear morphologic alterations resulting in cytostasis; (b) fiber diameter has little or no impact on in vitro toxicity when concentration is calculated as fibers/cm2; (c) fiber length is directly proportional to in vitro toxicity; and (d) toxicity of asbestos and vitreous fibers to CHO cells is not affected by composition. The lack of compositional effect in CHO cells does not correlate with findings from recent rodent inhalation studies using the same test fibers. Thus CHO cells may not be an appropriate in vitro model of fiber pathogenesis and would not constitute a valid toxicologic screening system for fibers.